Objective To investigate the effect of ursolic acid on the proliferation and apoptosis of human osteosarcoma cell line U2-OS and analyze its mechanism. Methods Human osteosarcoma cell line U2-OS was divided into 4 groups, which was cultured with ursolic acid of 0, 10, 20, and 40 μmol/L, respectively. At 0, 24, 48, and 72 hours after being cultured, the cell proliferation ability was detected by cell counting kit 8 (CCK-8). At 48 hours, the effects of ursolic acid on cell cycle and apoptosis of U2-OS cells were measured by flow cytometry. Besides, the expressions of cyclin D1 and Caspase-3 were detected by real-time fluorescent quantitative PCR and Western blot. Results CCK-8 tests showed that the absorbance (A) value of each group was not significant at 0 and 24 hours (P>0.05); but the differences between groups were significant at 48 and 72 hours (P<0.05). Flow cytometry results showed that, with the ursolic acid concentration increasing, the G1 phase of U2-OS cells increased, the S phase and G2/M phase decreased, and cell apoptosis rate increased gradually. There were significant differences between groups (P<0.05). Compared with the 0 μmol/L group, the relative expressions of cyclin D1 mRNA and protein in 10, 20, and 40 μmol/L groups significantly decreased (P<0.05); whereas, there was no significant difference in relative expression of Caspase-3 mRNA between groups (P>0.05). However, with the ursolic acid concentration increasing, the relative expressions of pro-Caspase-3 protein decreased and the relative expressions of activated Caspase-3 increased; there were significant differences between groups (P<0.05). Conclusion Ursolic acid can effectively inhibit the proliferation of osteosarcoma cell line U2-OS, induce the down-regulation of cyclin D1 expression leading to G0/G1 phase arrest, increase the activation of Caspase-3 and promote cell apoptosis.
ObjectiveTo investigate the expression of ErbB3 and Flotillin-2 in osteosarcoma biopsies and possible clinical pathology significance. MethodsThe tissue biopsies were harvested from 38 osteosarcoma patients and 13 osteochondroma patients between September 2009 and March 2014 for immunohistochemical staining. The ErbB3 and Flotillin-2 expressions were observed in osteosarcoma and osteochondroma biopsies, and the correlation and the relationship to the clinical and pathological features were analyzed. ResultsThe expression levels of ErbB3 and Flotillin-2 in osteosarcoma were significantly higher than those in osteochondroma (P<0.05). The high expressions of ErbB3 and Flotillin-2 had good consistency in osteosarcoma (Kappa=0.434, P=0.000). The high expression of ErbB3 was positively correlated to the clinical stages and lung metastasis (P<0.05), but it was not associated with gender, age, tumor location, and size (P>0.05). The high expression of Flotillin-2 had no correlation with clinical and pathological features (P>0.05). The influence factors of patients' overall survival included clinical stages, lung metastasis, high expression of ErbB3, and tumor size (P<0.05). Only lung metastasis and high expression of ErbB3 were independent factor affecting overall survival (P<0.05). ConclusionThe ErbB3 and Flotillin-2 express highly in osteosarcoma and the high expression has good consisitency. Besides, the high expression of ErbB3 is associated with the clinical stages and lung metastasis, indicating a poor prognosis for osteosarcoma.
目的 总结小细胞性骨肉瘤的临床特征及诊治情况。 方法 回顾性分析1995年12月－2012年6月收治的12例小细胞性骨肉瘤患者的临床资料。结合文献分析该病的诊治特点。 结果 全组患者手术11例，化学治疗（化疗）8例，术后辅助放射治疗（放疗）2例。随访截止时，全组患者总生存4～55个月。文献回顾方面，通过检索策略获得探讨相关治疗方案的文献8篇，涉及小细胞性骨肉瘤患者79例。治疗方式以手术+辅助化疗为主。 结论 小细胞性骨肉瘤发病率低，预后较普通骨肉瘤更差。手术为主要治疗方式，新辅助化疗及辅助化疗可能使患者进一步受益，放疗的治疗效果目前还不明确。
目的探讨甲状腺肉瘤的诊断与治疗。 方法回顾性分析2008年1月至2013年8月期间贵阳医学院附属医院甲状腺外科和贵州肿瘤医院甲状腺外科收治的5例甲状腺肉瘤患者的临床资料。 结果5例患者均行根治性手术，3例行术后化疗及放疗。术后均未发生声音嘶哑、呼吸困难、呛咳、肺部感染等并发症，切口均甲级愈合。经病理学免疫组化检查，具体分型为甲状腺组织细胞肉瘤1例，甲状腺血管肉瘤2例，甲状腺平滑肌肉瘤1例，甲状腺未分化肉瘤1例；波形蛋白（Vimentin）阳性5例，角蛋白（CK）阳性2例，平滑肌肌动蛋白（SMA）、结蛋白（Des）及S-100各阳性1例，甲状腺球蛋白（TG）、上皮膜抗原（EMA）、甲状腺转录因子（TTF）及降钙素均为阴性。术后5例患者均经电话随访，随访时间3～9个月，中位数为6个月。随访期间，3例患者因复发和转移而死亡；余2例仍在化疗及放疗中，无复发及转移。 结论甲状腺肉瘤的恶性程度高，宜采用以手术为主的综合治疗方案。
ObjectiveTo investigate the value of integrin αvβ3 targeted microPET/CT imaging with 68Ga-NODAGA-RGD2 as radiotracer for the detection of osteosarcoma and theranostics of osteosarcoma lung metastasis.MethodsThe 68Ga-NODAGA-RGD2 and 177Lu-NODAGA-RGD2 were prepared via one-step method and their stability and integrin αvβ3 binding specificity were investigated in vitro. Forty-one nude mice were injected with human MG63 osteosarcoma to established the animal model bearing subcutaneous osteosarcoma (n=21), osteosarcoma in tibia (n=5), and osteosarcoma pulmonary metastatic (n=15). The microPET-CT imaging was carried out in 3 animal models at 1 hour after tail vein injection of 68Ga-NODAGA-RGD2. Biodistribution study of 68Ga-NODAGA-RGD2 was performed in animal model bearing subcutaneous osteosarcoma at 10, 60, and 120 minutes. The animal model bearing pulmonary metastatic osteosarcoma was injected with 177Lu-NODAGA-RGD2 at 7 weeks after model establishment to observe the therapeutic effect of pulmonary metastatic osteosarcoma. Histological and immunohistochemistry examinations were also done to confirm the establishment of animal model and integrin β3 expression in animal models bearing subcutaneous osteosarcoma and bearing pulmonary metastatic osteosarcoma.Results68Ga-NODAGA-RGD2 and 177Lu-NODAGA-RGD2 had good stability in vitro with the 50% inhibitory concentration value of (5.0±1.1) and (6.5±0.8) nmol/L, respectively. The radiochemical purity of 68Ga-NODAGA-RGD2 at 1, 4, and 8 hours was 98.5%±0.3%, 98.3%±0.5%, and 97.9%±0.4%; while the radiochemical purity of 177Lu-NODAGA-RGD2 at 1, 7, and 14 days was 99.3%±0.7%, 98.7%±1.2%, and 96.0%±2.8%. 68Ga-NODAGA-RGD2 microPET-CT showed that the accumulation of 68Ga-NODAGA-RGD2 in animal models bearing subcutaneous osteosarcoma and osteosarcoma in tibia and in lung metastasis as small as 1-2 mm in diameter of animal model bearing pulmonary metastatic osteosarcoma. Biodistribution study of 68Ga-NODAGA-RGD2 in animal model bearing subcutaneous osteosarcoma revealed rapid clearance from blood with tumor peak uptake of (3.85±0.84) %ID/g at 120 minutes. The distribution of 177Lu-NODAGA-RGD2 in lung metastasis was similar with 68Ga-NODAGA-RGD2. The number and size of osteosarcoma metastasis decreased at 2 weeks after 177Lu-NODAGA-RGD2 administration and integrin targeting specificity was confirmed by pathology examination.Conclusion68Ga-NODAGA-RGD2 was potential for positive imaging and early detection of osteosarcoma and metastasis. Targeted radiotherapy with 177Lu-NODAGA-RGD2 was one potential alternative for osteosarcoma lung metastasis.
Objective To investigate early clinical manifestations of osteogenic sarcoma to help establishment of an early diagnosis of the disease.Methods A total of 92 patients with osteogenic sarcoma in the extremities were admitted to our hospital from April 1984 to October 2002. Of the 92 patients, 71 (42 males and 29 females; averaged age 17.4 years, range 666 years; illness course 1-28 weeks) had a complete record of their medical history and examination. From their first medical visits, we obtained their clinical symptoms, physical sings, diagnoses, and duration of the delayed diagnoses. The patients were pathologically confirmed as having osteogenic sarcoma in the extremities, with the lesions located in the distal femur in 38 patients, proximal tibia in 22, proximal femur in 3, proximal fibula in 3, proximal humerus in 2, distal tibia in 2, and distalradius in 1. Results Of the 71 patients, 70 had a local pain and/or a palpable mass, 37 had a persistent pain with no difference between day and night, 23 had an intermittent pain, and 11 had a nocturnal pain. Of the 71 patients, 42 had an initial pain related to trauma, and 3 of the 42 patients had a pathologic fracture. The patients with the local mass had a delayed diagnosis of osteogenic sarcoma with a delayed duration of 1-14 weeks, averaged 4 weeks; however, the patients without the local mass had a delayed diagnosis of this disease, with a delayed duration of 3-30 weeks averaged 14 weeks. In the patients undergoing an X-ray examination at the first medical visit, the duration of the delayed diagnoses was 1-20 weeks, averaged 8 weeks, but in the patients without an X-ray examination at first, the duration was 4-30 weeks, averaged 16 weeks. Conclusion Intermittent and persistent pains and local masses are the most characteristic clinical manifestations in the early stage of osteogenic sarcoma. A history of trauma often helps to make a diagnosis of the disease. Carefulclinical examination and observation should be given to adolescent patients whohave a recurrent pain around the joint.
Objective To investigate the possibility of using extendible distal femoral replacements in the treatment of osteosarcoma in growing individuals. Methods From December 1999 to March 2003, 3 cases (2 were typeⅡB, 1 was type ⅡA) with osteosarcoma were treated byextendible distal femoral replacements. Of the 3 cases, 2 underwent prosthesis extention operation, 1 was not operated. Results After the removal of tumor, the extremities of 2 patients were shortened by 4 to 5 cm within 2 to 3 years. After the lengthening procedure, the affected extremities were of equal length to the unaffected extremities and no drag symptoms of blood vessel and nerves were observed. Follow-up was done for 2 months to 3 years. There was no aseptic loosening. The function of joints was fairly good. Conclusion Extendible distal femoral replacements is an easy, convenient, and effective way to treat osteosarcoma.